The objective of this study was to compare the gait variability of patients with isolated anterior cruciate ligament (ACL) deficiency (experimental group) with that of healthy individuals (control group). The hypothesis was that the gait variability of the experimental group would be higher than the control group. The experimental group consisted of 20 men with an ACL tear and the control group consisted of 20 healthy men without any neurological and/or musculoskeletal pathology or injury. The gait acceleration signal was analysed using the Gait Evaluation Differential Entropy Method (GEDEM). The GEDEM index of the experimental group in the medio-lateral axis was significantly higher than that of the control subjects. A receiver operating characteristic (ROC) analysis was used to assess the diagnostic value of the method and to determine a cut-off entropy value. The GEDEM cut-off value had a 95.6% probability of separating isolated ACL patients from healthy subjects.